角质细胞叉头框蛋白O1基因调节血管内皮生长因子A的表达

发布时间：2018-04-11 15:55

本文选题：叉头框蛋白O + 血管内皮生长因子A　；参考：《第二军医大学学报》2017年03期

【摘要】：目的通过体外实验在角质细胞中过表达或沉默叉头框蛋白O1(FOXO1)基因,研究其对血管内皮生长因子A(VEGF-A)蛋白水平和转录活性的影响。方法采用体外细胞培养和转染方法,分别用ON-TARGET plus SMART pool人FOXO1siRNA和对照siRNA(ON-TARGET plus Non-targeting Control Pool)转染人永生化牙龈角质细胞(human immortalized gingival keratinocyte,HIGK),用免疫荧光法检测HIGK中FOXO1和VEGF-A的免疫荧光强度,荧光酶素实验分别检测FOXO1过表达和FOXO1沉默时VEGF-A的活性。结果与未转染组相比,使用FOXO1siRNA转染的HIGK中VEGF-A的蛋白水平降低了57%(P0.05)。荧光酶素实验分析结果显示,在HIGK中过表达FOXO1后VEGF-A的转录活性增加了2.1倍(P0.05),而FOXO1沉默时VEGF-A的转录活性减少了20%~43%(P0.05)。结论角质细胞中FOXO1参与VEGF-A的表达调控。
[Abstract]:Objective by in vitro experiments in keratinocytes overexpressing or silencing forkhead box protein O1 (FOXO1) gene on vascular endothelial growth factor A (VEGF-A) the effect of protein level and transcriptional activity. Methods using in vitro cell culture and transfection method, respectively using ON-TARGET plus SMART pool FOXO1siRNA siRNA (ON-TARGET plus Non-targeting and control Control Pool) transfected immortalized human gingival keratinocytes (human immortalized gingival keratinocyte, HIGK), the fluorescence intensity FOXO1 and VEGF-A detection in HIGK by immunofluorescence, luciferase experiment were used to detect the activity of VEGF-A FOXO1 expression and FOXO1 silencing. Results compared with untransfected group, VEGF-A protein levels using FOXO1siRNA transfection HIGK decreased by 57% (P0.05). Luciferase experiment results show that overexpression of FOXO1 VEGF-A transcription activity increased by 2.1 in HIGK (P0.05), while the transcriptional activity of VEGF-A decreased by 20%~43% (P0.05) when FOXO1 was silent. Conclusion FOXO1 was involved in the regulation of VEGF-A expression in keratinocytes.

【作者单位】：第二军医大学长海医院口腔科;上海交通大学医学院附属新华医院口腔科;
【基金】：上海交通大学医学院附属新华医院基金(134525)~~
【分类号】：R781.5

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